| BADH/pepB双价基因无选择标记表达载体转化苜蓿的初步研究 |
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| 引用本文： | 杨金慧,王丕武,曲静,王巍,王俊玲.BADH/pepB双价基因无选择标记表达载体转化苜蓿的初步研究[J].中国草地学报,2009,31(4). |
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| 作者姓名： | 杨金慧 王丕武 曲静 王巍 王俊玲 |
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| 作者单位： | 1. 吉林农业大学农学院,吉林,长春,130118
2. 吉林农业大学生命科学院,吉林,长春,130118 |
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| 基金项目： | 吉林省科技发展计划项目 |
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| 摘 要： | 选用公农1号紫花苜蓿,以5～7d莆龄的无菌子叶为外植体,通过农杆菌介导法将含有甜菜碱醛脱氢酶(BADH)和酸性蛋白酶(pepB)双价基因的无选择标记表达载体导入苜蓿子叶中,并用含有Na2CO3和NaHCO3碱性盐溶液的培养基进行筛选,得到抗盐碱转化植株.碱性盐浓度48mmol/L宜于子叶愈伤诱导,有利于转化植株的获得.对转化植株进行PCR检测,初步证明目的基囚序列已经整合剑苜楷基因组中.移栽成活的13棵碱性盐抗性植株经PCR检测有3棵植株呈阳性.
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| 关 键 词： | 无选择标记表达载体 转化 苜蓿 |
Research on Transforming BADH and pepB Bivalent Gene with Marker-free Expression Vector into Alfalfa |
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| YANG Jin-hui,WANG Pi-wu,QU Jing,WANG Wei,WANG Jun-ling.Research on Transforming BADH and pepB Bivalent Gene with Marker-free Expression Vector into Alfalfa[J].Chinese Journal of Grassland,2009,31(4). |
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| Authors: | YANG Jin-hui WANG Pi-wu QU Jing WANG Wei WANG Jun-ling |
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| Abstract: | The cotyledons of alfalfa were selected as explants for genetic transformation and were culti- vated with Agrobacterium tumefaciens strain EHA105 harboring marker-free expression vector containing BADH and pepB binary genes. Regenerated plants were obtained in the selection media supplemented with 48mmol/L Na2CO3 and NaHCO3. The genomic DNA of transgenic plants were extracted and analyzed by PCR. Result showed that BADH and pepB genes were transformed into the genome of transgenic alfalfa. 3 plants among 13 plants grown up through selection on medium containing 48mmol/L Na2CO3 and NaHCO3 were assessed by PCR amplification of 1340 bp of pepB gene. |
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| Keywords: | BADH pepB Marker-free expression BADH pepB Transformation Alfalfa Transgenic alfalfa |
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